Benzoylsulfonamides and sulfonylbenzamidines for use as antitumour agents

ABSTRACT

The present invention provides antitumor compounds of the formula (I), and antitumor methods

BACKGROUND OF THE INVENTION

In recent years fundamental advances have been made in the developmentof chemical agents and regimens of therapy to combat neoplasticdiseases. Despite these continuing advances, cancers continue to exactintolerable levels of human pain and suffering. The need for new andbetter methods of treating malignant neoplasms and leukemias continuesto fuel efforts to create new classes of compounds, especially in thearea of inoperable or metastatic solid tumors. The recent avalanche ofinformation regarding the basic biological processes involved inneoplasms has led to a deeper understanding of the heterogeneity oftumors. It is because of this extreme heterogeneity among populations ofneoplastic cells that new chemotherapeutic agents should have a widespectrum of activity and an acceptable therapeutic index. In addition,such agents must be chemically stable and compatible with other agents.It is also important that any chemotherapeutic regimen be as convenientand painless as possible to the patient.

Chemotherapy and radiation are frequently used in the treatment ofcancer and, although they often produce some response in the malignantdisease, they are rarely curative. Most solid tumors increase in massthrough the proliferation of malignant cells and stromal cells,including endothelial cells. In order for a tumor to grow larger than2–3 millimeters in diameter, it must form a vasculature, a process knownas angiogenesis. Suppression of tumor-induced angiogenesis byangiostatin and endostatin has been reported to result in antitumoractivity (O'Reilly, et al., Cell, 88, 277–285 (1997)). Becauseangiogenesis is a critical component of the mass expansion of most solidtumors, the development of new agents for the inhibition of this processrepresents a promising approach for antitumor therapy. This approach toantitumor therapy may lack the toxic side effects or drugresistance-inducing properties of conventional chemotherapy (JudahFolkman, Endoggnous Inhibitors of Angiogenesis, The Harvey Lectures,Series 92, pages 65–82, Wiley-Liss Inc., (1998)).

The N-[benzoyl]-phenylsulfonamides are well known in the agriculturalchemical arts as insecticides and herbicides (DE 2744137). The use ofN-[benzoyl]-phenylsulfonamides as antitumor agents generally, or asinhibitors of angiogenesis specifically, were heretofore notappreciated.

BRIEF SUMMARY OF THE INVENTION

The present invention provides a compound of Formula I:

where:

X is O or NH;

R¹ is hydrogen, halo, C₁–C₆ alkyl, C₁–C₄ alkoxy, C₁–C₄ alkylthio, CF₃,OCF₃, SCF₃, (C₁–C₄ alkoxy)carbonyl, nitro, azido, O(SO₂)CH₃, N(CH₃)₂,hydroxy, phenyl, substituted phenyl, pyridinyl, thienyl, furyl,quinolinyl, or triazolyl;

R² is hydrogen, halo, cyano, CF₃, C₁–C₆ alkyl, (C₁–C₄ alkoxy)carbonyl,C₁–C₄ alkoxy, phenyl, or quinolinyl;

R^(2a) is hydrogen or C₁–C₄ alkoxy;

R^(2b) is hydrogen or C₁–C₆ alkyl provided that at least one of R^(2a)and R^(2b) is hydrogen;

R³ is hydrogen, halo, C₁–C₆ alkyl, CF₃, or nitro;

R^(3a) is hydrogen, halo, or C₁–C₆ alkyl provided that when R^(3a) isC₁–C₆ alkyl, R³ is hydrogen and R⁴ is halo; and

R⁴ is halo, C₁–C₆ alkyl, or CF₃ provided that only one of R³ and R⁴ maybe C₁–C₆ alkyl and provided that when R⁴ is halo or C₁–C₆ alkyl only oneof R³ and R^(3a) is hydrogen; or a pharmaceutically acceptable baseaddition salt thereof, provided that:

-   -   a) when R³ and R⁴ are both chloro and R² is hydrogen, R¹ is        bromo, iodo, C₁–C₄ alkoxy, C₁–C₄ alkylthio, CF₃, OCF₃, nitro,        azido, O(SO₂)CH₃, N(CH₃)₂, hydroxy, phenyl, substituted phenyl,        pyridinyl, thienyl, furyl, or triazolyl;    -   b) when R³ and R⁴ are both chloro and R¹ is hydrogen, R² is        bromo, fluoro, CF₃, C₁–C₆ alkyl, C₁–C₄ alkoxy, phenyl, or        quinolinyl.

The present invention further provides a method of treating susceptibleneoplasms in a mammal comprising administering to a mammal in need ofsuch treatment an oncolytically effective amount of a compound ofFormula II:

where:

X is O or NH;

R¹ is hydrogen, halo, C₁–C₆ alkyl, C₁–C₄ alkoxy, C₁–C₄ alkylthio, CF₃,OCF₃, SCF₃, (C₁–C₄ alkoxy)carbonyl, nitro, azido, O(SO₂)CH₃, N(CH₃)₂,hydroxy, phenyl, substituted phenyl, pyridinyl, thienyl, furyl,quinolinyl, or triazolyl;

R² is hydrogen, halo, cyano, CF₃, C₁–C₆ alkyl, (C₁–C₄ alkoxy)carbonyl,C₁–C₄ alkoxy, phenyl, or quinolinyl;

R^(2a) is hydrogen or C₁–C₄ alkoxy;

R^(2b) is hydrogen or C₁–C₆ alkyl provided that at least one of R^(2a)and R^(2b) is hydrogen;

R³ is hydrogen, halo, C₁–C₆ alkyl, CF₃, or nitro;

R^(3a) is hydrogen, halo, or C₁–C₆ alkyl provided that when R^(3a) isC₁–C₆ alkyl, R³ is hydrogen and R⁴ is halo; and

R⁴ is halo, C₁–C₆ alkyl, or CF₃ provided that only one of R³ and R⁴ maybe C₁–C₆ alkyl and provided that when R⁴ is halo or C₁–C₆ alkyl only oneof R³ and R^(3a) is hydrogen; or a pharmaceutically acceptable baseaddition salt thereof.

The present invention also provides a method of suppressing tumorangiogenesis in a mammal comprising administering to a mammal in need ofsuch treatment an angiogenesis suppressing amount of a compound ofFormula II or a pharmaceutically acceptable base addition salt thereof.

The present invention also provides a pharmaceutical formulationcomprising a compound of Formula II or a pharmaceutically acceptablebase addition salt thereof, in combination with a pharmaceuticallyacceptable carrier, diluent or excipient.

This invention also provides the use of a compound of Formula II for themanufacture of a medicament for the treatment of susceptible neoplasms.Additionally, this invention provides a pharmaceutical formulationadapted for the treatment of susceptible neoplasms containing a compoundof Formula II. Furthermore, this invention includes a method for thetreatment of susceptible neoplasms that comprises administering aneffective amount of a compound of Formula II.

DETAILED DESCRIPTION OF THE INVENTION

The general chemical terms used in the formulae above have their usualmeanings. For example, the term “C₁–C₆ alkyl” includes methyl, ethyl,propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, andhexyl moieties. The term “C₁–C₄ alkyl” is included within the meaning ofC₁–C₆ alkyl and is taken to mean methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, and tert-butyl. The term “C₁–C₄ alkoxy” istaken to mean a C₁–C₄ alkyl group linked to the parent molecule throughan oxygen atom, and includes the groups methoxy, ethoxy, and isopropoxy.Likewise, the term “C₁–C₄ alkylthio” is taken to mean a C₁–C₄ alkylgroup linked to the parent molecule through a sulfur atom, and includesmethylthio, ethylthio, and isobutylthio. The term “halo” is taken tomean chloro, fluoro, bromo, and iodo. The term “substituted phenyl”means a mono-substituted phenyl wherein the substitutions are selectedfrom the group consisting of C₁–C₄ alkoxy, C₁–C₄ alkylthio, C₁–C₄ acyl,trifluoromethyl, and halo. The term “acyl” refers to an organic acidgroup in which the OH of the carboxy group is replaced by some othersubstituent (RCO—).

When X=NH, the molecule can exist in two tautomeric forms,

The present invention contemplates both of these forms.

While all of the compounds of Formula II are useful antitumor agents,certain classes of compounds are preferred. The following paragraphsdescribe such preferred classes.

-   -   a) R¹ is hydrogen and R² is bromo;    -   b) R¹ is fluoro and R² is chloro;    -   c) R¹ is fluoro;    -   d) R¹ is chloro;    -   e) R¹ is methyl;    -   f) R¹ is methylthio;    -   g) R² is hydrogen;    -   h) R³ is chloro, bromo, or CF₃;    -   i) R³ is chloro;    -   j) R³ is bromo;    -   k) R³ is CF₃;    -   l) R^(1a) is hydrogen;    -   m) R⁴ is chloro, bromo, methyl, or CF₃;    -   n) R⁴ is chloro;    -   o) R⁴ is bromo;    -   p) R⁴ is methyl;    -   q) R⁴ is CF₃;    -   r) R³ and R⁴ are both chloro;    -   s) R³ and R⁴ are both CF₃;    -   t) R³ is bromo and R⁴ is chloro;    -   u) R^(3a) is hydrogen and R³ and R⁴ are other than hydrogen;    -   v) X is O;    -   w) The compound of Formula II wherein the compound is a        pharmaceutically acceptable base addition salt;    -   x) The compound of Formula II wherein the compound is a sodium        salt;    -   y) R¹, R^(2a), and R^(2b) are hydrogen and R² is selected from        the group consisting of halo, C₁–C₄ alkyl, C₁–C₄ alkoxy, cyano,        trifluoromethyl, and quinolinyl;    -   z) R² and R^(2b) are hydrogen, R¹ is halo or C₁–C₄ alkyl, and        R^(2a) is C₁–C₄ alkyl or C₁–C₄ alkoxy; or    -   aa) R^(2a) is hydrogen, R¹ is C₁–C₄ alkoxy, and R² and R^(2b)        are C₁–C₄ alkyl.

Additionally, the following classes are especially preferred.

-   -   a) R², R^(2a), and R^(2b) are hydrogen and R¹ is selected from        the group consisting of hydrogen, halo, C₁–C₆ alkyl, C₁–C₄        alkoxy, C₁–C₄ alkylthio, CF₃, OCF₃, SCF₃, (C₁–C₄        alkoxy)carbonyl, nitro, azido, O(SO₂)CH₃, N(CH₃)₂, hydroxy,        phenyl, substituted phenyl, pyridinyl, thienyl, furyl,        quinolinyl, and triazolyl; or    -   b) R^(2a) and R^(2b) are hydrogen and R¹ is selection from the        group consisting of halo and C₁–C₄ alkyl, and R² is selected        from the group consisting of halo, C₁–C₄ alkyl, and C₁–C₄        alkoxycarbonyl.        It will be understood that the above preferred and especially        preferred classes may be combined to form additional preferred        and especially preferred classes.

The compounds of Formula II are antineoplastic agents. Thus, the presentinvention also provides a method of treating a susceptible neoplasm in amammal that comprises administering to a mammal in need of saidtreatment an oncolytically effective amount of a compound of Formula II.The present compounds are believed to be useful in treating carcinomassuch as neoplasms of the central nervous system: glioblastomamultiforme, astrocytoma, oligodendroglial tumors, ependymal and choroidplexus tumors, pineal tumors, neuronal tumors, medulloblastoma,schwannoma, meningioma, meningeal sarcoma; neoplasms of the eye: basalcell carcinoma, squamous cell carcinoma, melanoma, rhabdomyosarcoma,retinoblastoma; neoplasms of the endocrine glands: pituitary neoplasms,neoplasms of the thyroid, neoplasms of the adrenal cortex, neoplasms ofthe neuroendocrine system, neoplasms of the gastroenteropancreaticendocrine system, neoplasms of the gonads; neoplasms of the head andneck: head and neck cancer, oral cavity, pharynx, larynx, odontogenictumors; neoplasms of the thorax: large cell lung carcinoma, small celllung carcinoma, non-small cell lung carcinoma, malignant mesothelioma,thymomas, primary germ cell tumors of the thorax; neoplasms of thealimentary canal: neoplasms of the esophagus, neoplasms of the stomach,neoplasms of the liver, neoplasms of the gallbladder, neoplasms of theexocrine pancreas, neoplasms of the small intestine, veriform appendixand peritoneum, adneocarcinoma of the colon and rectum, neoplasms of theanus; neoplasms of the genitourinary tract: renal cell carcinoma,neoplasms of the renal pelvis and ureter, neoplasms of the bladder,neoplasms of the urethra, neoplasms of the prostate, neoplasms of thepenis, neoplasms of the testis; neoplasms of the female reproductiveorgans: neoplasms of the vulva and vagina, neoplasms of the cervix,addenocarcinoma of the uterine corpus, ovarian cancer, gynecologicsarcomas; neoplasms of the breast; neoplasms of the skin: basal cellcarcinoma, squamous cell carcinoma, dermatofibrosarcoma, Merkel celltumor; malignant melanoma; neoplasms of the bone and soft tissue:osteogenic sarcoma, malignant fibrous histiocytoma, chondrosarcoma,Ewing's sarcoma, primitive neuroectodermal tumor, angiosarcoma;neoplasms of the hematopoietic system: myelodysplastic sydromes, acutemyeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia,HILV-1 and T-cell leukemia/lymphoma, chronic lymphocytic leukemia, hairycell leukemia, Hodgkin's disease, non-Hodgkin's lymphomas, mast cellleukemia; and neoplasms of children: acute lymphoblastic leukemia, acutemyelocytic leukemias, neuroblastoma, bone tumors, rhabdomyosarcoma,lymphomas, renal tumors. In particular, the present compounds arebelieved to be useful in treating solid tumors, especially tumors of thecolon and rectum. It is preferred that the mammal to be treated by theadministration of the compounds of Formula II is human.

The compounds of the present invention are acidic in nature andaccordingly may react with any of a number of inorganic and organicbases, including amines and quaternary ammonium bases, to formpharmaceutically acceptable base addition salts. It is preferable toconvert the compounds of Formula II to their pharmaceutically acceptablebase addition salts for ease of administration when aqueous solutions ofthe subject compound are required. The Formula II compounds can reactwith basic materials such as alkali metal- or alkaline earth metalhydroxides, carbonates, and bicarbonates including, without limitation,sodium hydroxide, sodium carbonate, potassium hydroxide, calciumhydroxide, lithium hydroxide, etc. to form pharmaceutically acceptablesalts such as the corresponding sodium, potassium, lithium, or calciumsalt. The sodium and potassium salts are especially preferred.

Examples of amines suitable for forming salts are: primary, secondaryand tertiary aliphatic and aromatic amines, such as methylamine,ethylamine, propylamine, i-propylamine, the four isomeric butylamines,dimethylamine, diethylamine, diethanolamine, dipropylamine,diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine,trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine,quinoline and isoquinoline, especially ethyl-, propyl-, diethyl- ortriethylamine, but particulary isopropylamine and diethanolamine.

Examples of quaternary ammonium bases are in general the cations ofhaloammonium salts, for example the tetramethylammonium cation, thetrimethylbenzylammonium cation, the triethylbenzylammonium cation, thetetraethylammonium cation or the trimethylethylammonium cation, but alsothe ammonium cation.

The compounds of the present invention may be prepared by methods wellknown to one of ordinary skill in the art. Generally, theN-[benzoyl]-phenylsulfonamides of Formula II are prepared by coupling anappropriately substituted phenylsulfonamide with an appropriatelysubstituted benzoic acid or benzoic acid derivative as illustrated inthe following scheme. The variables R¹, R², R^(2a), R^(2b), R³, R^(3a),and R⁴ are as previously defined and Z is OH, Cl, Br,methanesulfonyloxy, or trifluoromethanesulfonyloxy.

When Z is OH, the corresponding benzoic acid is coupled to thephenylsulfonamide under standard peptide coupling conditions well knownto the skilled artisan. Specifically, the phenylsulfonamide and thebenzoic acid are coupled in the presence of a peptide coupling reagent,optionally in the presence of a catalyst. Suitable peptide couplingreagents include N,N′-carbonyldiimidazole (CDI),N,N′-dicyclohexylcarbodiimide (DCC),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), and1-(3-(1-pyrrolidinyl)propyl)-3-ethylcarbodiimide (PEPC). Polymersupported forms of EDC (Tetrahedron Letters, 34(48), 7685 (1993)) andPEPC (U.S. Pat. No. 5,792,763) have been described, and are very usefulfor the preparation of the compounds of the present invention. Suitablecatalysts for the coupling reaction include N,N-dimethyl-4-aminopyridine(DMAP). All of the reagents are combined in a suitable solvent,typically dichloromethane, chloroform, tetrahydrofuran, dioxane, ordiethyl ether and are stirred for from 1 to 72 hours at a temperature offrom ambient to about the reflux temperature of the solvent. The desiredproduct may be isolated by standard extractive and crystallizationtechniques, and purified by chromatography or crystallization asnecessary or desired. Where polymer-bound reagents are employed, theymay be conveniently removed from the reaction mixture by filtration.

Alternatively, the sulfonamide may be reacted with a benzoic acidderivative, such as compounds where Z is chloro, bromo,methanesulfonyloxy, or trifluoromethanesulfonyloxy, in the presence ofan acid scavenger such as pyridine, triethylamine, or a basic resin,optionally in the presence of a catalyst. The reagents are combined, andproducts isolated, essentially as described supra.

One skilled in the art would appreciate that compounds of Formula IIwhere X is NH may be prepared as illustrated in Scheme II where R¹, R²,R^(2a), R^(2b), R³, R^(3a), and R⁴ are as previously defined.

An appropriately substituted benzamidine is reacted with sulfonylderivatives, such as compounds where Z′ is chloro, bromo,methanesulfonyloxy, or trifluoromethanesulfonyloxy, in the presence ofan acid scavenger such as pyridine, triethylamine, or a basic resin,optionally in the presence of a catalyst. The reagents are combined, andproducts isolated, essentially as described supra.

The requisite benzoic acids, benzoic acid derivatives, benzamidines,sulfonyl derivatives and sulfonamides are either commercially availableor may be prepared by methods well known to the skilled artisan.

PREPARATION 1 2,4-dibromobenzonitrile

A solution of copper(I) cyanide (2.32 g, 25.9 mmol) in anhydrousdimethylsulfoxide (50 mL) is stirred at 60° C., and to this solution isadded tert-butylnitrite (7.1 mL, 59.7 mmol) all at once. A solution of2,4-dibromoaniline (5.0 g, 19.9 mmol) in anhydrous dimethylsulfoxide (30mL) is added dropwise, via cannula, to the mixture. After the additionis complete the reaction mixture is stirred for 1 hr, cooled to 45° C.,and then treated slowly with 5N HCl (50 mL). Five minutes later, thereaction mixture is cooled to ambient temperature and extracted with 1:1ethyl acetate:hexane (2×300 mL). The combined organic layers are washedwith water (100 mL) and saturated aqueous sodium chloride (100 mL),dried, concentrated under reduced pressure and the residue subjected tosilica gel chromatography, eluting with hexane containing from 0–5%ethyl acetate. Fractions containing product are combined andconcentrated under reduced pressure to provide the title compound (1.61g, 31% yield).

mp=76–78° C.

FDMS: m/e=261 (M⁺).

PREPARATION 2 2,4-dibromobenzoic acid

A stirred suspension of 2,4-dibromobenzonitrile (1.57 g, 6.0 mmol) insulfuric acid (6 M, 150 mL) is heated to reflux for 3 days. The reactionmixture is cooled down to ambient temperature and then extracted withethyl acetate (2×75 mL). The combined organic layers are washed withwater (100 mL) and saturated aqueous sodium chloride (50 mL), dried,concentrated, then subjected to silica gel chromatography, eluting withchloroform containing 0.5% methanol and 0.1% acetic acid. Fractionscontaining product are combined and concentrated under reduced pressureto provide the title compound (0.81 g, 48% yield).

mp=171–172° C.

ESIMS: m/e=279 (M⁺−1).

PREPARATION 3 2-bromo-chlorobenzoic acid

A solution of sodium nitrite (2.21 g) in water (15 mL) is added dropwiseto a stirred ice-cooled mixture of 2-amino-4-chlorobenzoic acid (5.00 g,29.1 mmol) and 48% hydrobromic acid (150 mL) in water (150 mL). Thereaction mixture is stirred for 2 hr at 0° C. and is then treateddropwise with a solution of copper(II) bromide (7.81 g) in water (20mL). Upon the completion of addition, the reaction mixture is allowed towarm to ambient temperature and is stirred overnight. The reactionmixture is then extracted with 3:1 ethyl acetate:hexane (2×400 mL). Thecombined organic layers are washed with saturated aqueous sodiumchloride (200 mL), dried, concentrated under reduced pressure, and theresidue subjected to silica gel chromatography, eluting with chloroformcontaining 1% methanol and 0.5% acetic acid. Fractions containingproduct are combined and concentrated under reduced pressure to providethe title compound (4.04 g, 59% yield).

mp=154–155° C.

ESIMS: m/e=233, 235 (M⁺−1).

PREPARATION 4 4-sulfamoylbenzoic acid methyl ester

4-Carboxyphenylsulfonamide (2.00 g, 9.9 mmol) is suspended in 3:1chloroform:methanol (200 mL). (Trimethylsilyl)-diazomethane is added asa 2.0 M solution in Hexanes (7.4 mL, 14.8 mmol) at ambient temperatureand stirred for 5 min. The solution is concentrated in vacuo, and thecrude is chromatographed on silica gel, 0.5% MeOH/0.1% AcOH in CH₂Cl₂.The product is a white solid, 2.11 g, 98% yield.

mp 180° C.

ESIMS m/e 214 (M⁺−1).

PREPARATION 5 3,4-dibromophenylsulfonamide

3,4-Dibromo-phenylsulfonyl chloride (20 mmol; Aldrich) is suspended in40 mL of 30% aqueous NHOH, and the mixture is stirred. Acetone is addedslowly, portionwise, to form a homogeneous reaction mixture (5–10 mL).This addition is exothermic, with vigorous bubbling. The reaction isstirred at room temperature and monitored by ESI-MS. The mixture isconcentrated by rotary evaporation to remove the acetone, and a solidformed. The solid is collected by suction filtration, washed with water,and allowed to air dry. The material is used as obtained, withoutfurther purification. ESIMS: 312, 314, 316 (M⁺−1); mp 169–171° C.; litmp 175–176° C. Huntress, E. H.: Carten, F. H. J. Am. Chem. Soc. 1940,62, 511–514.

The compounds of PREPARATION 6–15 are prepared essentially as describedin the procedure of PREPARATION 5.

PREP. Mass Spectral # Product Data (m/e) 6 3-chloro-4-fluoro- ESIMS:phenylsulfonamide 210 (M⁺ − 1) 7 4-iodo-phenylsulfonamide ESIMS: 284 (M⁺− 1) 8 3,4-dichlorophenylsulfonamide ESIMS m/e 224(M⁻ − 1; ³⁵Cl, ³⁵Cl)and 226 (M⁻ − 1; ³⁵Cl, ³⁷Cl) and 228 (M⁻ − 1; ³⁷Cl, ³⁷Cl) 94-isopropylphenylsulfonamide ESIMS m/e 198(M⁻ − 1) 104-ethylphenylsulfonamide ESIMS m/e 184(M⁻ − 1) 11 3-trifluoromethyl-ESIMS: 224 (M⁺ − 1) phenylsulfonamide 12 3-fluoro-phenylsulfonamideESIMS: 174 (M⁺ − 1) 13 toluene-3-sulfonamide ESIMS: 170 (M⁺ − 1) 143-bromophenylsulfonamide ESIMS: 237 (M⁺ − 1) 15 4-bromophenylsulfonamideESIMS: 237 (M⁺ − 1)

PREPARATION 16 2-chloro-4-methylbenzoic acid

To 4-bromo-3-chlorotoluene (4.97 g, 24.2 mmol) in DMF (25 mL) is addedPd(OAc)₂ (0.54 g, 2.42 mmol), 1,3-bis(diphenylphosphino)propane (0.998g, 2.42 mmol), triethylamine (12.5 mL) and methanol (12.5 mL). Thereaction vessel is evacuated and purged three times with carbon monoxidegas. A balloon filled with carbon monoxide gas is used to maintain thecarbon monoxide atmosphere. The reaction mixture is heated at 80° C. for8 hr. After cooling H₂O (50 mL) is added. The mixture is extracted withhexanes (2×50 mL). The combined organic layers are dried over Na₂SO₄,filtered, concentrated and chromatographed with 0–3% EtOAc in hexanes.1.24 g (28%) of methyl 2-chloro-4-methylbenzoate is isolated as acolorless oil. EIMS m/e 184 (M⁺; ³⁵Cl) and 186 (M⁺; ³⁷Cl).

To methyl 2-chloro-4-methylbenzoate (1.00 g, 5.42 mmol) in THF (10 mL)MeOH (5 mL) and H₂O (2.5 mL) is added 2N LiOH (8.12 mL, 16.2 mmol). Thereaction mixture is heated at 50° C. for 2.5 hr, cooled to roomtemperature, then quenched with 5N HCl (3.24 mL). The mixture isconcentrated to remove the TBF and MeOH. A white precipitate is formedand is filtered. After drying, 0.922 g (100%) of2-chloro-4-methylbenzoic acid is isolated. ESIMS m/e 169 (M⁻−1; ³⁵Cl)and 171 (M⁻−1; ³⁷Cl).

PREPARATION 17 4-(tert-butyldimethylsilyloxy)phenylsulfonamide

4-Hydroxyphenylsulfonamide (3.46 g, 20 mmol) is dissolved in DMF (40 mL)and treated with tert-butyl-dimethylsilylchloride (3.31 g, 22.0 mmol)and imidazole (1.50 g, 22.0 mmol) at room temperature. After 20 hr, thereaction mixture is diluted with EtOAc (100 mL) and washed with 1.0 NHCl (2×50 mL). The organic phase is dried (MgSO4), filtered, andconcentrated to yield an oil. The crude oil is purified by Biotagecolumn chromatography (40M SiO₂ column, eluted at 75 mL/min with 1:1hexanes:EtOAc). A white solid is obtained (4.24 g, 15.4 mmol, 77%).ESI-MS m/e 288.1 (M⁺+H); mp 117–118° C.; ¹H NMR (CDCl₃) δ 7.78 (d, 2H),6.89 (d, 2H), 4.86 (br s, 2H), 0.97 (s, 9H), 0.20 (s, 6H).

PREPARATION 18N-(2,4-dichlorobenzoyl)-4-(tert-butyldimethylsilyloxy)-phenylsulfonamide

To a stirring solution of 2,4-dichlorobenzoic acid (1.25 eq) in drydichloromethane (10 mL/mmol),4-(tert-butyl-dimethylsiloxy)phenylsulfonamide (1.0 eq) is added in oneportion followed by EDC (1.25–1.5 eq) and finally,NN-dimethyl-4-aminopyridine (1.2 equiv). The mixture is vigorouslystirred under nitrogen for 16 hr, concentrated under reduced pressure,and the residue partitioned between ethyl acetate and water. The organiclayer is washed with 1N hydrochloric acid (4 times, 20 mL/mmol), thenthe combined aqueous phases extracted with ethyl acetate (twice, 20mL/mmol). The combined organic layers are finally washed with water andsaturated aqueous sodium chloride, dried over sodium sulfate andconcentrated under reduced pressure. The residue is purified by silicagel chromatography or crystallization if necessary or desired. ESI-MSm/e 458.0 (M⁺−H; 460.0 (M⁺+H).

The compounds of PREPARATION 19–21 are prepared essentially as describedin the procedure of PREPARATION 18.

PREP. Mass Spectral # Product Data (m/e) 19 3-Bromo-N-(2,4-dichloro-ESI-MS m/e benzoyl)-phenylsulfonamide 409.0(M⁺ − H) 204-Iodo-N-(2,4-dichloro- ESI-MS m/e benzoyl)-phenylsulfonamide 455.08(M⁺− H) 21 4-Bromo-N-(2,4-dichloro- ESI-MS m/e benzoyl)-phenylsulfonamide409.0(M⁺ − H)

General Coupling Procedure

To a stirring solution of the benzoic acid (1.25 eq) in drydichloromethane (10 mL/mmol), the phenylsulfonamide (1.0 eq) is added inone portion followed by EDC (1.25–1.5 eq) and finally,N,N-[dimethyl]4-aminopyridine (1.2 equiv). The mixture is vigorouslystirred under nitrogen for 16 hr, concentrated under reduced pressure,and the residue partitioned between ethyl acetate and water. The organiclayer is washed with 1N hydrochloric acid (4 times, 20 mL/mmol), thenthe combined aqueous phases extracted with ethyl acetate (twice, 20mL/mmol). The combined organic layers are finally washed with water andsaturated aqueous sodium chloride, dried over sodium sulfate andconcentrated under reduced pressure. The residue may be subjected tosilica gel chromatography or crystallization if necessary or desired.

The compounds of EXAMPLES 1–84 are prepared essentially as described inthis general procedure.

EXAM- PLE Mass Spectral # Product Data (m/e) 1 N-[2,4- MS(ES):dichlorobenzoyl]-3- 408 [M − H]⁻ bromophenylsulfonamide 2 N-[2,4-MS(ES): dichlorobenzoyl]-3- 362 [M − H]⁻ chlorophenylsulfonamide 3N-[2,4- MS(ES): dichlorobenzoyl]-4- 360 [M − H]⁻methoxyphenylsulfonamide 4 N-[2,4- ESIMS: dibromobenzoyl]-4- 434 (M⁺ ⁺¹⁾ methylphenylsulfonamide 5 N-[2,4- ESIMS: dibromobenzoyl]- 476 (M⁺ ⁺¹⁾ 4-tert- butylphenylsulfonamide 6 N-[2,4- ESIMS: dibromobenzoyl]-4-454, 456(M⁺ + 1) chlorophenylsulfonamide 7 N-[2-bromo-4- ESIMS:chlorobenzoyl]-4- 388, 390(M⁺ + 1) methylphenylsulfonamide 8N-[2-bromo-4- ESIMS: chlorobenzoyl]-4- 408, 410(M⁺ + 1)chlorophenylsulfonamide 9 N-[2,4- ESIMS: dichlorobenzoyl]-3- 380(M⁺ −2), chloro-4- 382(M⁺), fluorophenylsulfonamide 384(M⁺ + 2) 10N-[2-chloro-4- ESIMS: nitrobenzoyl]-4- 373(M⁺ − 2),chlorophenylsulfonamide 375(M⁺), 377(M⁺ + 2) 11 N-[2-chloro-4- ESIMS:bromobenzoyl]- 372(M⁺ − 2), phenylsulfonamide 374(M⁺) 12 N-[2-methyl-4-ESIMS: bromobenzoyl]- 352(M⁺ − 2), phenylsulfonamide 354(M⁺) 13N-[2-chloro-4- ESIMS: nitrobenzoyl]- 339(M^(+ − 1),) phenylsulfonamide341(M⁺ + 1) 14 N-[2-chloro-4- ESIMS: bromobenzoyl]-4- 450(M⁺ − 3),bromophenylsulfonamide 452(M⁺ − 1), 454(M⁺ + 1) 15 N-[2-chloro-4- ESIMS:nitrobenzoyl]-4- 417(M⁺ − 2), bromophenylsulfonamide 419(M⁺) 16N-[2-chloro-4- ESIMS: bromobenzoyl]-4- 390(M⁺ − 2),fluorophenylsulfonamide 392(M⁺) 17 N-[2-chloro-4- ESIMS:bromobenzoyl]-3- 406(M⁺ − 3), chlorophenylsulfonamide 408(M⁺ − 1),410(M⁺ + 1) 18 N-[2-chloro-4- ESIMS: bromobenzoyl]-4- 402(M⁺ − 2),methoxyphenylsulfonamide 404(M⁺), 406(M⁺ + 2) 19 N-[2-methyl-4- ESIMS:bromobenzoyl]-4- 382(M⁺ − 2), methoxyphenylsulfonamide 384(M⁺) 20N-[2-chloro-4- ESIMS: nitrobenzoyl]-4- 369(M⁺ − 1),methoxyphenylsulfonamide 371(M⁺) 21 N-[2-chloro-4- ESIMS:nitrobenzoyl]-3,4- 407(M⁺ − 2), dichlorophenyl- 409(M⁺), sulfonamide411(M⁺ + 2) 22 N-[2-methyl-4- ESIMS: chlorobenzoyl]-3- 342(M⁻ − 1),chlorophenylsulfonamide 344(M⁻ − 1), 346(M⁻ − 1) 23 N-[2-methyl-4- ESNegative Ion MS chlorobenzoyl]-4- [M − H]⁻ methylphenylsulfonamide ionsobserved: m/z 342(³⁵Cl, ³⁵Cl), m/z 344 (³⁵Cl, ³⁷Cl) and m/z 346 (³⁷Cl,³⁷Cl). 24 N-[2,4- ES Negative Ion MS dichlorobenzoyl]-3,4- [M − H]⁻dibromophenylsulfonamide ions observed: m/z 322 (³⁵Cl) and m/z 324(³⁷Cl). 25 N-[2,4- ESIMS: dichlorobenzoyl]-3- 395.9476/395.9469trifluoromethyl- phenylsulfonamide 26 N-[2,4- ESIMS: dichlorobenzoyl]-3-345.9508/345.9515 fluorophenylsulfonamide 27 N-[2,4- ESIMS:dichlorobenzoyl]-3- 365.9734/365.9747 methylphenylsulfonamide 28N-[2-methyl-4- ESIMS m/e 386.0, bromobenzoyl]-4- 387.9, and 389.9chlorophenylsulfonamide (M⁺ − 1; ⁷⁹Br, ³⁵Cl; ⁷⁹Br, ³⁷Cl; ⁸¹Br, ³⁷Cl) 29N-[2-methyl-4- ESIMS m/e 366.0 and bromobenzoyl]-4- 368.0 (M⁺ − 1;methylphenylsulfonamide ⁷⁹Br; ⁸¹Br) 30 N-[2-bromo-4- ESIMS m/e 430.0,431.9 chlorobenzoyl]-4- and 433.9 (M⁺ − 1; methyoxycarbonyl- ³⁵Cl, ⁷⁹Br;³⁷Cl, phenylsulfonamide ⁷⁹Br; ³⁷Cl, ⁸¹Br) 31 N-[2,4- ESIMS m/e 473.8,dibromobenzoyl]-4- 475.9, and 477.9 methoxycarbonyl- (M^(+ − 1;)phenylsulfonamide ⁷⁹Br, ⁷⁹Br; ⁷⁹Br, ⁸¹Br; ⁸¹Br, ⁸¹Br) 32 N-[2-bromo-4-ESIMS m/e 459.9, chlorobenzoyl]-3- 461.9, 463.9 methoxycarbonyl-4- (M⁺ −1; methoxyphenylsulfonamide ⁷⁹Br, ³⁵Cl; ⁸¹Br, ³⁵Cl; ⁸¹Br, ³⁷Cl) 33N-[2-bromo-4- ESIMS m/e 428.2, chlorobenzoyl]- 430.2, and 432.2; 4-tert-(M^(+ − 1;) butylphenylsulfonamide ⁷⁹Br, ³⁵Cl; ⁸¹Br, ³⁵Cl; ⁸¹Br, ³⁷Cl 34N-[2,4- ESIMS m/e 503.9, dibromobenzoyl]-3- 506.0 and 508.0;methoxycarbonyl-4- (M⁺ − 1; methoxyphenylsulfonamide ⁷⁹Br, ⁷⁹Br, ⁸¹Br,⁷⁹Br; ⁸¹Br, ⁸¹Br) 35 N-[2-methyl-4- ESIMS m/e 404.0, bromobenzoyl]-3-406.0, 408.0 chloro-4- (M⁺ − 1; fluorophenylsulfonamide ⁷⁹Br, ³⁵Cl;⁸¹Br, ³⁵Cl; ⁸¹Br, ³⁷Cl) 36 N-[2-methyl-4- ESIMS m/e 478.0 andbromobenzoyl]-4- 480.0 (M⁺ − 1; iodophenylsulfonamide ⁷⁹Br; ⁸¹Br) 37N-[2-methyl-4- ESIMS m/e 366.1 and bromobenzoyl]-3- 368.1 (M⁺ − 1;methylphenylsulfonamide ⁷⁹Br; ⁸¹Br) 38 N-[2,4- ESIMS m/e 541.9,dibromobenzoyl]-4- 543.9 and 545.9 iodophenylsulfonamide (M^(+ − 1;)⁷⁹Br, ⁷⁹Br; ⁸¹Br, ⁷⁹Br; ⁸¹Br, ⁸¹Br) 39 N-[2,4- ESIMS m/e 454dichlorobenzoyl]-4- (M⁻ − 1; iodophenylsulfonamide ³⁵Cl, ³⁵Cl) and 456(M⁻ − 1; ³⁵Cl, ³⁷Cl) and 458 (M⁻ − 1; ³⁷Cl, ³⁷Cl) 40 N-[2-chloro-4- ESNegative Ion MS bromobenzoyl]-4- [M − H]⁻ iodophenylsulfonamide ionsobserved: m/z 498 (⁷⁹Br, ³⁵Cl), m/z 500 (⁸¹Br, ³⁵Cl) and m/z 502 (⁸¹Br,³⁷Cl). 41 N-[2-methyl-4- ES Negative Ion MS chlorobenzoyl]-4- [M − H]⁻methoxyphenylsulfonamide ions observed: m/z 338 (³⁵Cl) and m/z 340(³⁷Cl). 42 N-[2-methyl-4- ES Negative Ion MS chlorobenzoyl]-4- [M − H]⁻fluorophenylsulfonamide ions observed: m/z 326 (³⁵Cl) and m/z 328(³⁷Cl). 43 N-[2-methyl-4- ES Negative Ion MS chlorobenzoyl]-4- [M − H]⁻bromophenylsulfonamide ions observed: m/z 386 (⁷⁹Br, ³⁵Cl), m/z 388(⁸¹Br, ³⁵Cl) and m/z 390 (⁸¹Br, ³⁷Cl). 44 N-[2-methyl-4- ES Negative IonMS chlorobenzoyl]-2- [M − H]⁻ methoxy-4- ions observed:methylphenylsulfonamide m/z 352 (³⁵Cl) and m/z 354 (³⁷Cl). 45N-[2-methyl-4- ES Negative Ion MS chlorobenzoyl]-3- [M − H]⁻ chloro-4-ions observed: methylphenylsulfonamide m/z 356(³⁵Cl, ³⁵Cl), m/z 358(³⁵Cl, ³⁷Cl) and m/z 360 (³⁷Cl, ³⁷Cl). 46 N-[2,4-bis- ES Negative Ion MStrifluoromethylbenzoyl]- [M − H]⁻ 4- ion observed:fluorophenylsulfonamide m/z 414. 47 N-[2,4-bis- ES Negative Ion MStrifluoromethylbenzoyl]- [M − H]⁻ 4- ion observed:methoxyphenylsulfonamide m/z 426. 48 N-[2,4-bis- ES Negative Ion MStrifluoromethylbenzoyl]- [M − H]⁻ 4- ion observed:methylphenylsulfonamide m/z 410. 49 N-[2,4-bis- ES Negative Ion MStrifluoromethylbenzoyl]- [M − H]⁻ 4- ions observed:bromophenylsulfonamide m/z 474 (⁷⁹Br) and m/z 476 (⁸¹Br). 50 N-[2,4-bis-ES Negative Ion MS trifluoromethylbenzoyl]- [M − H]⁻ 3- ion observed:methylphenylsulfonamide m/z 410. 51 N-[2-methyl-4- ES Negative Ion MSchlorobenzoyl]-4- [M − H]⁻ trifluoromethoxy- ions observed:phenylsulfonamide m/z 392 (³⁵Cl) and m/z 394 (³⁷Cl). 52 N-[2,4-bis- ESNegative Ion MS trifluoromethylbenzoyl]- [M − H]⁻ 3,4-dichlorophenyl-ions observed: sulfonamide m/z 464 (³⁵Cl, ³⁵Cl), m/z 466 (³⁵Cl, ³⁷Cl)and m/z 468 (³⁷Cl, ³⁷Cl). 53 N-[2,4-bis- ES Negative Ion MStrifluoromethylbenzoyl]- [M − H]⁻ 3,4-difluorophenyl- ion observed:sulfonamide m/z 432. 54 N-[2-methyl-4- ES Negative Ion MSchlorobenzoyl]-3- [M − H]⁻ chloro-4- ions observed:fluorophenylsulfonamide m/z 360 (³⁵Cl, ³⁵Cl), m/z 362 (³⁵Cl, ³⁷Cl) andm/z 364 (³⁷Cl, ³⁷Cl). 55 N-[2-methyl-4- ES Negative Ion MSchlorobenzoyl]-3- [M − H]⁻ methylphenylsulfonamide ions observed: m/z322 (³⁵Cl) and m/z 324 (³⁷Cl). 56 N-[2-methyl-4- ES Negative Ion MSchlorobenzoyl]-4- [M − H]⁻ iodophenylsulfonamide ions observed: m/z 434(³⁵Cl) and m/z 436 (³⁷Cl). 57 N-[2-methyl-4- ES Negative Ion MSchlorobenzoyl]-3, [M − H]⁻ 4-difluorophenyl- ions observed: sulfonamidem/z 344 (³⁵Cl) and m/z 346 (³⁷Cl). 58 N-[2,4-bis- ES Negative Ion MStrifluoromethylbenzoyl]- [M − H]⁻ 3-chloro-4- ions observed:fluorophenyl- m/z 448 (³⁵Cl) and sulfonamide m/z 450 (³⁷Cl). 59N-[2-chloro-4- ES Positive Ion MS methylbenzoyl]- [M + H]⁺ 3-chloro-4-ions observed: fluorophenyl- m/z 362 (³⁵Cl) and sulfonamide m/z 364(³⁷Cl). 60 N-[2-methyl-4- ES Positive Ion MS chlorobenzoyl]- [M + H]⁺phenylsulfonamide ions observed: m/z 310 (³⁵Cl) and m/z 312 (³⁷Cl). 61N-[2-methyl-4- ES Negative Ion MS bromobenzoyl]-4- [M − H]⁻ethylthiophenyl- ions observed: sulfonamide m/z 412 (⁷⁹Br) and m/z 414(⁸¹Br). 62 N-[2,4- ES Negative Ion MS dichlorobenzoyl]-4- [M − H]⁻ethylthiophenyl- ions observed: sulfonamide m/z 388 (³⁵Cl, ³⁵Cl), m/z390 (³⁵Cl, ³⁷Cl) and m/z 392 (³⁷Cl, ³⁷Cl). 63 N-[2,4-bis- ES NegativeIon MS trifluoromethylbenzoyl]- [M − H]⁻ 4-isopropylphenyl- ionobserved: sulfonamide m/z 438. 64 N-[2-chloro-4- ES Negative Ion MSbromobenzoyl]-4- [M − H]⁻ isopropylphenyl- ions observed: sulfonamidem/z 414 (⁷⁹Br, ³⁵Cl), m/z 416 (⁸¹Br, ³⁵Cl) and m/z 418 (⁸¹Br, ³⁷Cl). 65N-[2,4- ES Negative Ion MS dibromobenzoyl]-4- [M − H]⁻ethylphenylsulfonamide ions observed: m/z 444 (⁷⁹Br, ⁷⁹Br), m/z 446(⁷⁹Br, ⁸¹Br) and m/z 448 (⁸¹Br, ⁸¹Br). 66 N-[2-methyl-4- ES Negative IonMS bromobenzoyl]-4- [M − H]⁻ ethylphenylsulfonamide ions observed: m/z380 (⁷⁹Br) and m/z 382 (⁸¹Br). 67 N-[2,4- ES Positive Ion MSdibromobenzoyl]-4- [M + H]⁺ isopropylphenyl- ions observed: sulfonamidem/z 460 (⁷⁹Br, ⁷⁹Br), m/z 462 (⁷⁹Br, ⁸¹Br) and m/z 464 (⁸¹Br, ⁸¹Br). 68N-[2,4- ES Negative Ion MS dichlorobenzoyl]-4- [M − H]⁻ isopropylphenyl-ions observed: sulfonamide m/z 370 (³⁵Cl, ³⁵Cl), m/z 372 (³⁵Cl, ³⁷Cl)and m/z 374 (³⁷Cl, ³⁷Cl). 69 N-[2,4- ES Negative Ion MSdichlorobenzoyl]-4- [M − H]⁻ ethylphenylsulfonamide ions observed: m/z356 (³⁵Cl, ³⁵Cl), m/z 358 (³⁵Cl, ³⁷Cl) and m/z 360 (³⁷Cl, ³⁷Cl). 70N-[2-methyl-4- ES Negative Ion MS chlorobenzoyl]-4- [M − H]⁻ethylphenylsulfonamide ions observed: m/z 336 (³⁵Cl) and m/z 338 (³⁷Cl).71 N-[2-chloro-4- ES Negative Ion MS bromobenzoyl]-4- [M − H]⁻ethylphenylsulfonamide ions observed: m/z 400 (⁷⁹Br, ³⁵Cl), m/z 402(⁸¹Br, ³⁵Cl) and m/z 404 (⁸¹Br, ³⁷Cl). 72 N-[2-bromo-4- ES Negative IonMS chlorobenzoyl]-4- [M − H]⁻ ethylphenylsulfonamide ions observed: m/z400 (⁷⁹Br, ³⁵Cl), m/z 402 (⁸¹Br, ³⁵Cl) and m/z 404 (⁸¹Br, ³⁷Cl). 73N-[2-bromo-4- ES Negative Ion MS chlorobenzoyl]-4- [M − H]⁻isopropylphenyl- ions observed: sulfonamide m/z 414 (⁷⁹Br, ³⁵Cl), m/z416 (⁸¹Br, ³⁵Cl) and m/z 418 (⁸¹Br, ³⁷Cl). 74 N-[2-chloro-4- ESIMS m/e460.9. iodobenzoyl]-4- (M − 1) azidophenylsulfonamide 75 N-[2-methyl-4-ES Positive Ion MS chlorobenzoyl]-4- [M + H]⁺ methylthiophenyl- ionsobserved: sulfonamide m/z 356 (³⁵Cl) and m/z 358 (³⁷Cl). 76N-[2-methyl-4- ES Negative Ion MS chlorobenzoyl]-4- [M − H]⁻trifluoromethylthio- ions observed: phenylsulfonamide m/z 408 (⁷⁹Br,³⁵Cl), m/z 410 (⁸¹Br, ³⁵Cl) and m/z 412 (⁸¹Br, ³⁷Cl). 77 N-[2-methyl-4-ES Positive Ion MS chlorobenzoyl]-3- [M + H]⁺ trifluoromethyl- ionsobserved: phenylsulfonamide m/z 378 (³⁵Cl) and m/z 380 (³⁷Cl). 78N-[2-methyl-4- ES Positive Ion MS bromobenzoyl]-3- [M + H]⁺trifluoromethyl- ions observed: phenylsulfonamide m/z 422 (⁷⁹Br) and m/z424 (⁸¹Br). 79 N-[2-trifluoromethyl- ES Positive Ion MS4-methylbenzoyl]- [M + H]⁺ 4-methylphenyl- ion observed: sulfonamide m/z358. 80 N-[2,4- ¹H NMR (CDCl3) δ dichlorobenzoyl]- 7.68–7.153-methoxyphenyl- (m, 7 H); sulfonamide 3.80 (s, 3 H). 81 N-[2,4- ESI-MSm/e 395.9 dichlorobenzoyl]- (M⁺ − H) 4-trifluoromethyl-phenylsulfonamide 82 N-[2,4- Mp 155–157° C.; dichlorobenzoyl]- ESI-MSm/e 344.0 4-hydroxyphenyl- (M⁺ − H) sulfonamide 83 N-[4-Bromo-2- MS(ES): methyl-benzoyl]- [M − H]⁻ 3,4-dimethyl- 379.9943 phenylsulfonamide84 N-[2,4-Bis- MS (ES): trifluoromethyl- [M − H]⁻ benzoyl]-3-fluoro-414.0049 phenylsulfonamide

EXAMPLE 85 N-[2-chloro-4-bromobenzoyl]-4-chlorophenylsulfonamide

An 8 mL reaction vial is charged with 2-chloro-4-bromobenzoic acid (0.39mmol, 1.5 eq) and 2.0 mL of dichloromethane. A stock solution (4.0 mL)containing 4-chlorophenylsulfonamide (0.26 mmol, 1 eq) andN,N-[dimethyl]-4-aminopyridine (48 mg, 0.39 mmol, 1.5 eq) indichloromethane is added, followed by 0.261 g carbodiimide polystyreneresin (2.0 mmol/g, 0.52 mmol, 2.0 eq, Novabiochem) and the vial iscapped and rotated. After 72 hr, 0.77 g sulphonated polystyrene resin(NP-TsOH) is added (1.53 mmol/g, 1.17 mmol, Argonaut). After about 18 hrthe reaction mixture is filtered and concentrated under a stream ofnitrogen. The residue is subjected to reverse phase HPLC; CombiPrepcolumn, YMC ODS-A 20×50 mm column with 5 micron, C18, 120 Angstrom poresize, gradient: 5% to 95% CH₃CN/0.01 HCl aqueous solution. Fractionscontaining product are combined and concentrated under reduced pressureto provide the title compound.

ESIMS: m/e=408(M⁺+1), 406(M⁺−1), 410(1⁺+3).

The compounds of Examples 86–107 are prepared essentially as describedin Example 85.

EXAM- PLE Mass Spectral # Product Data (m/e) 86N-[2,4-dichlorobenzoyl]-4- ESIMS: methylphenylsulfonamide 342(M⁺ − 1),344(M⁺ + 1) 87 N-[2,4-dichlorobenzoyl]-4- ESIMS:methylthiophenylsulfonamide 374(M⁺ − 1), 376(M⁺ + 1) 88N-[2,4-dichlorobenzoyl]-4- ESIMS: tert- 384(M⁺ − 1),butylphenylsulfonamide 386(M⁺ + 1) 89 N-[2,4-dichlorobenzoyl]-3- ESIMS:chloro-4- 378(M⁺ + 1), methylphenylsulfonamide 376(M⁺ − 1), 380(M⁺ + 3)90 N-[2-methyl-4-chlorobenzoyl]-3- ESIMS: bromophenylsulfonamide388(M⁺ + 1), 386(M⁺ − 1), 390(M⁺ + 3) 91 N-[2,4-dichlorobenzoyl]-4-ESIMS: fluorophenylsulfonamide 346(M⁺ − 1), 348(M⁺ + 1) 92N-[2,4-dichlorobenzoyl]-3,4- ESIMS: dichlorophenylsulfonamide 398(M⁺ +1), 396(M⁺ − 1), 400(M⁺ + 3) 93 N-[2-methyl-4-chlorobenzoyl]-4- ESIMS:chlorophenylsulfonamide 342(M⁺ − 1), 344(M⁺ + 1) 94N-[2,4-dichlorobenzoyl]-4- ESIMS: bromo 408(M + 1), phenylsulfonamide406(M − 1), 410(M + 3) 95 N-[2,4-dichlorobenzoyl]-4- ESIMS:methylsulfonyloxyphenyl- 422 (M − 2), sulfonamide 424 (M), 426 (M + 2)96 N-[2,4-dichlorobenzoyl]-4- ESIMS: trifluoromethoxyphenyl- 412 (M −2), sulfonamide 414 (M), 416 (M + 2) 97 N-[2,4-dichlorobenzoyl]-4-ESIMS: methoxy-3,5- 368 (M − 2), dimethylphenylsulfonamide 370 (M), 372(M + 2) 98 N-[2,4-dichlorobenzoyl]-4- ESIMS: dimethylaminophenyl- 371 (M− 2), sulfonamide 373 (M) 99 N-[2,4-bis- ESIMS:trifluoromethylbenzoyl]-4- 430 (M − 1), chlorophenylsulfonamide 432(M + 1) 100 N-[2-methyl-4-bromobenzoyl]-4- ESIMS: bromophenylsulfonamide430 (M − 3), 432 (M − 1), 434 (M + 1) 101 N-[2-chloro-4-nitrobenzoyl]-4-ESIMS: fluoro-phenylsulfonamide 357 (M − 1) 102N-[2-chloro-3-methylbenzoyl]-4- ESIMS: chlorophenylsulfonamide 342 (M− 1) 344 (M + 1) 103 N-[2,4-dichlorobenzoyl]-4- ESIMS:nitrophenylsulfonamide 373 (M − 1) 375 (M + 1) 104N-[2-methyl-4-chlorobenzoyl]-3,4- ESIMS: dichlorophenylsulfonamide 378(M + 1) 376 (M − 1) 380 (M + 3) 105 N-[2-methyl-4-chlorobenzoyl]-4-ESIMS: tert-butylphenylsulfonamide 364 (M − 1) 366 (M + 1) 106N-[2-methyl-4-chlorobenzoyl]-3- ESIMS: cyanophenylsulfonamide 333 (M− 1) 107 N-[2,4-dichlorobenzoyl]- 395 (M) 4-[1,2,4]triazol-4-yl-phenylsulfonamide

EXAMPLE 108N-[2,4-dichlorobenzoyl]-4-(1-methylsulfanylo-phen-4-yl)phenylsulfonamide

Step A: Procedure for Activation of the Resin

The Rink amide resin (CA Novabiochem, 0.53 mmol/g) was suspended in a30% solution of pyridine in DMF and stirred at room temperature for 3hours. The mixture was filtered and the resin was washed twice with DMFand then, alternatively with CH₂Cl₂ and MeOH. The activated resin havinga free amino group was dried and used without further purification.

The Rink amide resin (0.53 mmol/g) was suspended in a 1:1 mixtureCH₂Cl₂/THF and Et₃N (4 eq), 4-iodophenylsulfonamide (3 eq) and DMAP(catalytic amount). The solution was stirred overnight at roomtemperature. The mixture was filtered and the resin was washedalternatively with CH₂Cl₂ and MeOH. The 4-iodophenylsulfonamide Rinkresin was dried under vacuum.

The corresponding 4-iodophenylsulfonarmide Rink resin (0.26 mmol, 0.53mmol/g), Methylsulfanyl-phenyl boronic acid (2 eq), potassium carbonate(6 eq) and the Palladium acetate (0.5 eq) were mixed together andsuspended in 7 mL of a mixture dioxane/water 6:1. This mixture washeated in an Argovant® QUEST® 210 at 100° C. for 24 hours. Then, theresin was washed twice with 5 mL of a mixture dioxane/water 6:1 and thensix times with CH₂Cl₂ (7 nL) followed each time by MeOH (7 mL).

3 mL of a 95% aqueous solution of trifluoroacetic acid were added to theresin previously dissolved in 3 mL of CH₂Cl₂. The mixture was stirredfor 30 min at room temperature, filtered as described above. The4′-methylsulfanyl-biphenylsulfonamide was employed without furtherpurification.

To a stirred solution of 2,4-Dichloro-benzoic acid (1.25 eq) in dryCH₂Cl₂ (10 ml/mmol), 4′-Methylsulfanyl-biphenyl-4-sulfdnamide (1.0 eq)was added in one portion followed by EDC (1.25 or 1.5 eq) and finally,DMAP (1.2 equiv). The mixture was vigorously stirred under nitrogen for16 hours, then evaporated in vacuo and the residue partitioned betweenEtOAc and water. The organic layer was washed with 1N HCl (4 times, 20mL/mmol), then the aqueous phase was extracted with EtOAc (twice, 20mL/mmol). The combined organic layers were finally washed with water andbrine, dried over Na₂SO₄ and concentrated in vacuo. The crude productwas purify by silica gel chromatography using the appropriate eluentafford the title compound.

ESI-MS (M⁺−H) 450.9870/450.0.

EXAMPLE 109 N-[2,4-dichlorobenzoyl]-4-3′-acetyl-biphenylsulfonamide

A suspension of 4-iodophenylsulfonamide Rink resin (0.26 mmol, 0.53mmol/g), 3-acetylphenyl boronic acid (2 eq) and 2,4-dichloro-benzoicacid (1.25 eq); were used essentially as described in Example 108 toprepare the title compound.

ESI-MS (M⁺−H) 447.0099/446.0.

EXAMPLE 110 N-[2,4-dichlorobenzoyl]phenylsulfonamide

To a mixture of phenylsulfonamide (0.16 mol; 25.12 g) and potassiumcarbonate (0.2 mol; 27.6 g) in 500 mL dioxane is added dropwise2,4-dichlorobenzoyl chloride (0.13 mole; 18.0 mL). The mixture is warmedto reflux under nitrogen for 16 hr. The reaction is then diluted withwater (500 mL), neutralized to pH 5 with concentrated hydrochloric acid,and extracted 3 times with ethyl acetate. The combined ethyl acetatelayers are washed with saturated aqueous sodium chloride, dried oversodium sulfate and concentrated under reduced pressure to a white solid.The solid residue is subjected to silica gel chromatography, elutingwith dichloromethane containing from 0–5% methanol. Fractions containingthe product are combined and concentrated under reduced pressure toprovide the title compound.

MS(ES): m/e=329.9 (M⁺+1), 327.9 (M⁺−1).

EXAMPLE 111 N-[2,4-dichlorobenzoyl]-4-chlorophenylsulfonamide

A mixture of 4-chlorophenylsulfonamide (0.1 mol; 19.0 g) and2,4-dichlorobenzoyl chloride (0.12 mol; 16.8 mL); the title compound wasprepared essentially as described in Example 110.

MS(ES): m/e=363.9 (M+)

EA: Calculated for C₁₃H₈Cl₃NO₃S: Theory: C, 42.82; H, 2.21; N, 3.84.Found: C, 42.56; H, 2.14; N, 3.76.

EXAMPLE 112 25 N-[2-chloro-4-bromobenzoyl]-4-chlorophenylsulfonamide

To a reaction mixture of 4-chlorophenylsulfonamide (15.6 g, 81.4 mmol),CDI (15.82 g, 97.7 mmol) and ethyl acetate (300 mL) at room temperatureis added a slurry of 2-chloro-4-bromobenzoic acid (23.0 g, 97.7 mmol) inethyl acetate (100.0 mL) over a period of 15.0 min (note: gas evolutionis observed which can be controlled by the rate of addition of theslurry; reaction mixture goes into solution by the end of addition ofthe slurry; reaction can be monitored by HPLC or TLC with 1:1 ethylacetate/heptane eluent. The reaction is then stirred at room temperaturefor 30 min and then heated at 60° C. for 90 min or until no gasevolution is observed. The reaction is cooled to 40° C., and1,8-diazabicyclo[5.4.0]undec-7-ene is (14.63 mL) added (all at once).The reaction temperature goes from 40° C. to 45° C. The mixture isstirred until it reaches room temperature before quenching withdeionized water (400 mL). The top organic layer is separated, washedwith 1N HCl (300.0 mL), dried with anhydrous MgSO₄, filtered and thecake washed with ethyl acetate (20.0 mL). The filtrate is concentratedto 50.0 g of syrupy solution, then heptane (250.0 mL) is added withvigorous stirring. With heating, a white slurry is formed and isrefluxed and then allowed to equilibrate to room temperature. The whiteprecipitate is filtered and the cake is washed with heptane (20.0 mL).The precipitate is dried in a house vacuum at 55° C. for 18 hr.(mass=29.12 g, 87.4% wt yield).

A mixture of 19.17 g of the product and 1:2 ethyl acetate/heptane (150.0mL) is heated at reflux for 30 min, and then cooled to room temperature.The off white precipitate is filtered and then the cake is washed withheptane (50.0 mL). The precipitate is dried in a house vacuum at 50° C.for 18 hr. (mass=14.93 g; 78% recovery).

ESIMS: m/e=408(M⁺+1),406(M⁺−1), 410(M⁺+3).

EXAMPLE 113 N-[2-chloro-4-bromobenzoyl]-4-chlorophenylsulfonamide sodiumsalt

To a solution of N-[2-chlorobromobenzoyl]-4-chlorophenylsulfonamide (5.2g, 12.72 mmol) and tert-butyl methyl ether (88.0 mL) at room temperatureis added sodium methoxide (0.69 g, 12.72 mmol) all at once. The reactionis then stirred for 5 hr, after which heptane (88.0 mL) is addedfollowed by vigorous stirring for 60 min. A white precipitate is formed,filtered off under a positive nitrogen pressure, and the cakesubsequently washed with heptane (2×44.0 mL). The cake is dried tosemi-dryness, followed by drying in a house vacuum oven at 130° C. for18 hr (mass=4.4 g, 80% wt. Yield; ¹H nmr (DMSO d₆) 7.8–7.85 (m, 1H),7.81–7.82 (m, 1H), 7.58–7.59 (d, 1H, J=1.76 Hz), 7.51–7.52 (m, 1H),7.48–7.49 (m, 1H), 7.44–7.45 (d, 1H, J=1.76) 7.37–7.4 (d, 1H).

EXAMPLE 114N-[3-chloro-4-fluorophenylsulfonyl]-3-fluoro-4-methylbenzamidine

Add 3-fluoro-4-methylbenzamidine hydrochloride (0.025 g, 0.133 mmol) inTHF (0.5 mL) to 3-chloro-4-fluorophenyl sulfonylchloride (0.0304 g,0.133 mmol) followed by N-methylmorpholine (0.2 mL). The reactionmixture was concentrated and chromatographed using reverse phasechromatography (gradient of 5–95% (0.1% TFA in CH3CN) in (0.1% TFA inH₂O). A white solid (16.4 mg, 36%) was isolated. ES Positive Ion MS[M+H]⁺ ions observed: m/z 345 (³⁵Cl) and m/z 347 (³⁷Cl).

EXAMPLE 115 N-[3-chloro-4-fluorophenylsulfonyl]-4-chlorobenzamidine

4-chlorobenzamidine hydrochloride (0.025 g, 0.133 mmol) and 3-chloro-4fluorophenyl sulfonylchloride (0.0304 g, 0.133 mmol); were usedessentially as described in Example 114 to prepare the title compound.ES Positive Ion MS [M+H]⁺ ions observed: m/z 347 (³⁵Cl, ³⁵Cl), m/z 349(³⁷Cl, ³⁷Cl) and m/z 351 (³⁷Cl, ³⁷Cl).

EXAMPLE 116N-[3-chloro-4-fluorophenylsulfonyl]-3-chloro-4-fluorobenzamidine

A mixture of 3-chloro-4-fluorobenzamidine hydrochloride (0.025 g, 0.133mmol) and 3-chloro-4-fluorophenyl sulfonylchloride (0.0304 g, 0.133mmol); the title compound is prepared essentially as described inExample 115. ES Positive Ion MS [M+H]⁺ ions observed: m/z 365 (³⁵Cl,³⁵Cl), m/z 367 (³⁵Cl, ³⁷Cl) and m/z 369 (³⁷Cl, ³⁷Cl).

EXAMPLE 117 25 N-[2,4-dichlorobenzoyl]-4-hydroxyphenylsulfonamide

4-Methoxy-phenyl-4-sulfonamide (0.0608 g, 0.132 mmol) is dissolved inTHF (1.25 mL) and treated with tetrabutylammonium fluoride (1.0 N/THF;200 μL, 2.0 mmol) at room temperature with stirring for 18 hr. Thereaction mixture is diluted with EtOAc (10 mL) and washed with saturatedaq. NH₄Cl (1 mL), H₂O (2×1 nm), and brine (1 mL). The organic phase isdried MgSO4, filtered, and concentrated by rotary evaporation.(Lyopholized from H₂O/MeOH to obtain a glassy solid, 20 mg (0.058 mmol,58%). Purified by preparative HPLC.) mp 155–157° C.; ESI-MS m/e 344.0(M+−H); ¹H NMR (d6-DMSO) 7.90 (d, 2H); 7.68 (s, 1H); 7.44 (s, 2H); 6.90(d, 2H); 3.43 (br s, 3H).

EXAMPLE 118 N-[2,4-dichlorobenzoyl]-4-(thien-3-yl)-phenylsulfonamide

To a solution of N-(2,4-dichlorobenzoyl)-4-iodo-phenylsulfonamide (0.10mmol) in toluene/ethanol 20/1 (3 mL) is added 3-thiopheneboronic acid(0.18 mmol, 0.18 mL, 1.0M solution in DMF) andtetrakis-(triphenylphosphine) palladium (0) (10 mol %). Then 2M aqueousNa₂CO₃ is added (0.3 mL) and the stirred mixture is heated to 100° C.overnight (17 hr)(Buchi Syncore system). The reaction mixture isconcentrated (Genevac apparatus), then water is added (2.5 mL) and ethylacetate (5 mL). The phases are separated and the aqueous layer isextracted with ethyl acetate (3×5 mL). This process is automaticallycarried out using a Tecan system. The solvents are evaporated and thecorresponding crude product is purified by HPLC to give the titlecompound.

ESI-MS m/e 410.96/410.0 [M+−H]

The compounds of EXAMPLES 119–130 are prepared essentially as describedin the procedure for EXAMPLE 118.

EXAM- PLE Mass Spectral # Product Data (m/e) 119N-[2,4-dichlorobenzoyl]-4- ESI-MS m/e (1-trifluoromethylphen-4-yl)472.99/472.2 phenylsulfonamide [M⁺ − H] 120 N-[2,4-dichlorobenzoyl]-4-ESI-MS m/e (1-fluoro-phen-3-yl)- 422.99/422.0 phenylsulfonamide [M⁺ − H]121 N-[2,4-dichloro-benzoyl]-4- ESI-MS m/e furan-2-yl-phenylsulfonamide394.98/394.2 [M⁺ − H] 122 N-[2,4-dichlorobenzoyl]-4- ESI-MS m/e(1-methoxy-phen-2-yl) 435.01/434.0 phenylsulfonamide [M⁺ − H] 123N-[2,4-dichlorobenzoyl]-4- ESI-MS m/e (1-methoxycarbonyl-phen-447.07/446.0 4-yl)phenylsulfonamide [M⁺ − H] 124N-[2,4-dichlorobenzoyl]-4- ESI-MS m/e quinolin-8-yl-phenylsulfonamide456.01/455.1 [M⁺ − H] 125 N-[2,4-dichloro-benzoyl]-3- ESI-MS m/equinolin-8-yl-phenylsulfonamide 456.01/457.2 [M⁺ + H] 126N-[2,4-dichlorobenzoyl]-4- ESI-MS m/e fur-3-yl-phenylsulfonamide394.98/394.0 [M⁺ − H] 127 N-[2,4-dichlorobenzoyl]-4- ESI-MS m/epyridin-3-yl-phenylsulfonamide 406.0 [M⁻ − H] 128N-[2,4-dichloro-benzoyl]-3- ESI-MS m/e biphenylsulfonamide 405.1 [M⁻ −H] 129 N-[2,4-dichloro-benzoyl]-4′- ESI-MS m/emethoxy-4-biphenylsulfonamide 435.2 [M⁻ − H] 130N-[2,4-dichlorobenzoyl]-4- ESI-MS m/e thiophen-2-yl-phenylsulfonamide411.1 [M⁻ − H]

All of the compounds concerned are orally available and are normallyadministered orally, and so oral administration is preferred. However,oral administration is not the only route or even the only preferredroute. For example, transdermal administration may be very desirable forpatients who are forgetful or petulant about taking oral medicine, andthe intravenous route may be preferred as a matter of convenience or toavoid potential complications related to oral administration. Compoundsof Formula II may also be administered by the percutaneous,intramuscular, intranasal or intrarectal route in particularcircumstances. The route of administration may be varied in any way,limited by the physical properties of the drugs, the convenience of thepatient and the caregiver, and other relevant circumstances (Remington'sPharmaceutical Sciences, 18th Edition, Mack Publishing Co. (1990)).

The pharmaceutical compositions are prepared in a manner well known inthe pharmaceutical art. The carrier or excipient may be a solid,semi-solid, or liquid material that can serve as a vehicle or medium forthe active ingredient. Suitable carriers or excipients are well known inthe art. The pharmaceutical composition may be adapted for oral,inhalation, parenteral, or topical use and may be administered to thepatient in the form of tablets, capsules, aerosols, inhalants,suppositories, solutions, suspensions, or the like.

The compounds of the present invention may be administered orally, forexample, with an inert diluent for capsules or compressed into tablets.For the purpose of oral therapeutic administration, the compounds may beincorporated with excipients and used in the form of tablets, troches,capsules, elixirs, suspensions, syrups, wafers, chewing gums and thelike. These preparations should contain at least 4% of the compound ofthe present invention, the active ingredient, but may be varieddepending upon the particular form and may conveniently be between 4% toabout 70% of the weight of the unit. The amount of the compound presentin compositions is such that a suitable dosage will be obtained.Preferred compositions and preparations of the present invention may bedetermined by methods well known to the skilled artisan.

The tablets, pills, capsules, troches, and the like may also contain oneor more of the following adjuvants: binders such as povidone,hydroxypropyl cellulose, microcrystalline cellulose, or gelatin;excipients or diluents such as: starch, lactose, microcrystallinecellulose or dicalcium phosphate; disintegrating agents such as:croscarmellose, crospovidone, sodium starch glycolate, corn starch andthe like; lubricants such as: magnesium stearate, steric acid, talc orhydrogenated vegetable oil; glidants such as colloidal silicon dioxide;wetting agents such as: sodium lauryl sulfate and polysorbate 80 (CASNo. 9005-65-6); and sweetening agents such as: sucrose, aspartame orsaccharin may be added or a flavoring agent such as: peppermint, methylsalicylate or orange flavoring. When the dosage unit form is a capsule,it may contain, in addition to materials of the above type, a liquidcarrier such as polyethylene glycol or a fatty oil. Other dosage unitforms may contain other various materials that modify the physical formof the dosage unit, for example, as coatings. Thus, tablets or pills maybe coated with sugar, hydroxypropyl methylcellulose, polymethacrylates,or other coating agents. Syrups may contain, in addition to the presentcompounds, sucrose as a sweetening agent and certain preservatives, dyesand colorings and flavors. Materials used in preparing these variouscompositions should be pharmaceutically pure and non-toxic in theamounts used.

Injections for parenteral administration include sterile aqueous ornon-aqueous solutions, suspensions and emulsions. Aqueous solutions andsuspensions may include distilled water for injection or physiologicalsalt solution. Non-aqueous solutions and suspensions may includepropylene glycol, polyethylene glycol, vegetable oil such as olive oil,alcohol such as ethanol or polysorbate 80. Injections may compriseadditional ingredients other than inert diluents: e.g. preservingagents, wetting agents, emulsifying agents, dispersing agents,stabilizing agents (such as lactose), assisting agents such as agents toassist dissolution (e.g. glutamic acid or aspartic acid). They may besterilized for example, by filtration through a bacteria-retainingfilter, by incorporation of sterilizing agents in the compositions or byirradiation. They may also be manufactured in the form of sterile solidcompositions which may be dissolved in sterile water or some othersterile diluent(s) for injection immediately before use.

The compounds of Formula II are generally effective over a wide dosagerange. For example, dosages per day normally fall within the range ofabout 10 to about 300 mg/kg of body weight. In some instances dosagelevels below the lower limit of the aforesaid range may be more thanadequate, while in other cases still larger doses may be employedwithout causing any harmful side effect, and therefore the above dosagerange is not intended to limit the scope of the invention in any way.

It will be understood that the amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound or compoundsadministered, the age, weight, and response of the individual patient,and the severity of the patient's symptoms.

Inhibition of HUVEC Proliferation

Human umbilical vein endothelial cells (JC; BioWhittaker/Clonetics,Walkersville, Md.) were maintained in endothelial cell growth medium(EGM) containing basal medium (EBM) with bovine brain extract, humanepidermal growth factor, hydrocortisone, gentamicin, amphotericin B and2% fetal bovine serum. For the assay, HUVEC (5×10³) in EBM (200 μl) with0.5% fetal bovine serum were added to wells in a 96-well cell cultureplate and incubated at 37° C. for 24 hr in humidified 5% carbondioxide/air. The test compounds were serially diluted in dimethylsulfoxide (DMSO) in concentrations from 0.0013 to 40 μM and added to thewells in 20 μl. Then human vascular endothelial growth factor (VEGF) (20ng/mL in wells; R&D Systems, Minneapolis, Minn.) prepared from a stocksolution of 100 μg/mL in phosphate buffered normal saline containing0.1% bovine serum albumin, was added to the wells. The HUVEC wereincubated at 37° C. for 72 hr in humidified 5% carbon dioxide/air. WST-1cell proliferation reagent (20 μL; Boehringer Mannheim, Indianapolis,Ind.) was added to the wells and the plates returned to the incubatorfor 1 hr. The absorbance of each well at 440 nm was measured. The growthfraction was determined from the absorbance of treated wells with andwithout VEGF divided by the absorbance obtained from control wells setto zero and 1.0. The exemplified compounds were tested in this assay andall exhibited an IC₅₀≦1.0 μM.

Rat Corneal Micropocket Assay

Fisher 344 female rats (145–155 grams; Taconic, Inc., Germantown, N.Y.)were anesthesized with acepromazine (2.5 mg/kg, ip) 20 minutes prior toinitiation of 2–3% isoflurane/oxygen inhalation. The body temperaturewas maintained with a circulating hot water pad. The surgery wasperformed using an ophthalmic operating microscope (OMS.75 OperatingMicroscope, TopCon Corporation, Japan). A scalpel blade (#15) was usedto make a vertical half-thickness linear corneal incision just lateralto the center of the eye. The tip of the scalpel blade was used togently undermine the superior corneal layer of the cornea nearest to thelimbus. A pocket was formed in the cornea using blunt dissection withcorneal scissors (Roboz, Rockville, Md.). Nitrocellulose filters (0.45μM, Millipore, Bedford, Mass.) were cut into small disks using a 20gauge needle punch. The disks were soaked in 2 μl of human VEGF solution(0.82 μg/l; R&D Systems) or human basic fibroblast growth factor (0.20μg/μl; R&D Systems) for 10 minutes on ice. Using forceps, the disksimpregnated with the angiogenic factor (VEGF or bFGF) were inserted intothe corneal pocket so that the disk is firmly covered with cornealepithelium. The animals were treated with the compound of Example 110(160 mg/kg) administered orally by gavage in phosphate buffered salineonce per day on days 1 through 10 post implantation of the disks. Theeyes were photographed on days 7 and 14 post implantation of the disks.For photography, the animals were treated with atropine sulfate (AmTechGroup, Inc., Phoenix Scientific, Inc., St. Joseph, Mo.) topically formydriasis and anesthetized with 2–3% isoflurane/oxygen. The eyes werephotographed using the ophthalmic microscope and the images were savedusing Image Pro-Plus software. The images were analyzed by convertingthe area of interest to high contrast black and white reversed image andcounting the bright pixels as a determination of the vascular area. Thedata are images from at least 6 eyes. The compound of Example 110 was avery effective inhibitor of VEGF-induced neoangiogenesis, but was not aneffective inhibitor of bFGF-induced neoangiogenesis.

HCT116 Colon Carcinoma Cell Growth Inhibition

HCT116 colon carcinoma cells were grown monolayer culture in RPMI 1640medium supplemented with 10% fetal bovine serum and 1%penicillin-streptomycin (GibcoBRL, Grand Island, N.Y.). HCT116 cells inexponential growth phase were exposed to various concentrations of thetest compounds at 37° C. for 72 hr in 5% carbon dioxide/air. Afterexposure to the agent, the cells were washed with 0.9% phosphatebuffered saline. Growth inhibition was determined using WST-1 cellproliferation reagent as described above. The results are expressed asthe growth fraction of treated cells compared with control cultures.Representative compounds of the present invention were tested forefficacy against the human colon HCT116 tumor cells. The data from theseexperiments are summarized in TABLE I.

TABLE I IC₅₀ IC₅₀ EXAMPLE (μM) EXAMPLE (μM) 1 4.2 62 19.3 3 2.5 63 24.75 3.8 64 18.2 6 4.0 70 16.9 9 1.7 73 8.3 10 12.1 79 5.2 11 5.5 80 12.314 40.0 81 11.3 15 10.4 82 12.4 16 3.8 86 1.6 17 15.8 87 5.5 18 9.3 884.8 20 2.4 89 4.8 21 30.4 90 4.5 23 15.9 92 17.4 24 37.1 93 18.1 25 9.694 3.6 26 6.1 95 4.3 30 10.1 96 34.7 32 7.8 97 7.0 33 7.8 98 13.0 3416.5 99 33.5 35 17.0 100 9.2 36 28.4 101 12.4 37 15.5 102 6.3 38 13.9103 11.9 41 14.5 104 22.2 42 3.3 105 14.8 44 18.7 106 4.6 45 20.7 10816.0 46 4.8 109 25.1 47 8.7 110 3.5 49 14.7 111 15.0 50 10.5 112 2.8 5126.7 115 33.6 53 11.2 116 5.9 55 9.5 117 12.4 56 16.8 121 24.5 57 5.0122 25.8 59 15.0 123 37.7 60 6.6 124 5.6 61 14.2 126 18.5Conventional Murine Tumor and Human Tumor Xenouraft Assays

Inhibition of tumors transplanted into mice is an accepted procedure forstudying the efficacy of antitumor agents (Corbett, et al., In vivoMethods for Screening and Preclinical Testing; Use of rodent solidtumors for drug discovery., In: Anticancer Drug Development Guide:Preclinical Screening, Clinical Trials, and Approval, B. Teicher (ed),Humana Press Inc., Totowa, N.J., Chapter 5, pages 75–99 (1997);(Corbett, et al., Int., J. Parmacog., 33, Supplement, 102–122 (1995)).Murine tumors or human xenographs were implanted essentially asdescribed by Corbett in In vivo Methods for Screening and PreclimicalTesting; Use of rodent solid tumors for drug discovery. Briefly, themurine tumor or human xenograph was implanted subcutaneously usingeither 12-gauge trocar implants or counted number of cells. The locationfor the trocar insertion is midway between the axillary and inguinalregion along the side of the mouse. The trocar is slipped approximately¾ of an inch subcutaneously up toward the axilla before discharging thetumor fragment, and pinching the skin as the trocar is removed.,Alternatively, human tumor cells prepared from a brie of donor tumors(5×10⁶ cells) were implanted subcutaneously in a hind-leg of a male orfemale nude mouse (Charles River). Either a test compound in vehicle orvehicle alone was administered by intravenous bolus injection (iv),intraperitoneal injection (ip), or oral gavage (po). Each treatmentgroup, as well as a group of untreated control animals, consisted offive animals per group in each experiment. Subcutaneous tumor responsewas monitored by tumor volume measurement performed twice each week overthe course of the experiment (60–120 days). Body weights were taken as ageneral measure of toxicity. The subcutaneous tumor data were analyzedby determining the median tumor weight for each treatment group over thecourse of the experiment and calculating the tumor growth delay as thedifference in days for the treatment versus the control tumors to reacha volume of either 500 or 1000 mm³.

The compound of Example 110 was tested against a variety of murine andhuman tumors substantially as described supra. The data from these testsare summarized in TABLES II–XIII. The parameters measured in eachexperiment are summarized in the following paragraphs.

-   -   Tumor Weight(mg)=(a×b²)/2 where a=tumor length (mm) and b=tumor        width (mm).    -   Tumor Growth Delay=T−C where T is the median time (days)        required for the treatment group tumors to reach a predetermined        size, and C is the median time (days) for the control group        tumors to reach the same size. Tumor-free survivors are excluded        from this calculation, and are tabulated separately (Tumor        Free).

${{Log}\mspace{14mu}{Kill}} = \frac{{Tumor}\mspace{14mu}{Growth}\mspace{14mu}{Delay}}{(3.32)({Td})}$

-   -   where Tumor Growth Delay is as previously defined and Td is        tumor volume doubling time (days), estimated from the best fit        straight line from a log-linear growth plot of the control group        of tumors in exponential growth (100–800 mg range)    -   % T/C mass—The treatment and control groups are measured when        the control group tumors reach approximately 700 to 1200 mg in        size (median group). The median tumor weight of each group is        determined (including zeros). The T/C value in percent is an        indication of antitumor effectiveness. A T/C≦42% is considered        significant antitumor activity. A T/C<10% is considered to        indicate highly significant antitumor activity.    -   Body Weight Loss Nadir—A body weight loss nadir (mean of group)        of greater than 20% or drug deaths greater than 20% are        considered to indicate an excessively toxic dosage in single        course trials.    -   Activity Rating—the Activity Rating is derived from the Log Kill        according to the following table:

ANTITUMOR ACTIVITY ACTIVITY LOG KILL RATING HIGHLY ACTIVE >2.8 ++++2.0–2.8 +++ 1.3–1.9 ++ 0.7–1.2 + INACTIVE <0.7 −

TABLE II EARLY STAGE MURINE COLON ADENOCARCINOMA-38 (BDF1 male mice,CRL-Raleigh) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAY FREEAT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 144RATING   0 +6.3% 0/5 — — — 0/5 − 1200^(a) −5.2% 0/6 0% All cures >4.56/6 ++++  920^(b) +7.7% 0/5 0% All cures >4.5 5/5 ++++  272^(c) +3.1%0/5 0% All cures >4.5 5/5 ++++ ^(a)Drug was administered IV at 80 mg/kgon day 3; 160 mg/kg on day 4; 240 mg/kg on day 5; and 120mg/kg/injection BID on days 7–9. ^(b)Drug was administered IV at 40mg/kg on day 3; 80 mg/kg on day 4; and 160 mg/kg/injection QD on days5–9. ^(c)Drug was administered IV at 20 mg/kg on day 3; 40 mg/kg on day4; and 106 mg/kg/injection QD on days 5–6.

TABLE III EARLY STAGE MURINE MAMMARY ADENOCARCINOMA-16/C (C3H femalemice, CRL-Kingston) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAYFREE AT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 14RATING  0 +0.0% 0/5 — — — 0/5 − DILUENT +10.8 0/4 60% 0.7 0.22 0/4 −INFUSION 560^(a) DEATH 5/5 — DEATH — 0/5 DEATH 560^(b) +10.9% 0/4 37%2.0 0.6 0/4 + 680^(c) +7.6% 0/5 39% 2.5 0.75 0/5 + ^(a)Drag wasadministered at 560 mg/kg by IV infusion over 3 hours on day 3. ^(b)Drugwas administered at 280 mg/kg/by IV infusion on days 3 and 7. ^(c)Drugwas administered IV at 80 mg/kg/injection on days 3–6; 120mg/kg/injection on days 7–9.

TABLE IV EARLY STAGE HUMAN COLON ADENOCARCINOMA-HCT116 (Balb/C SCIDfemale mice, NCI) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAYFREE AT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 115RATING  0 −4.2% — — — — 0/5 − 560^(a) −4.0% 0/5 0% 56 6.7 2/5 ++++280^(b)    0% 0/5 0% 28 3.4 2/5 ++++ ^(a)Drug was administered 140mg/kg/injection IV on days 3, 4, 5, and 9. ^(b)Drug was administered 70mg/kg/injection IV on days 3, 4, 5, and 9.

TABLE V EARLY STAGE TAXOL SENSITIVE HUMAN COLON ADENOCARCINOMA #15/0(Balb/C SCID female mice, NCI) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS ATDRUG DELAY FREE AT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOGKILL DAY 151 RATING  0 +4.3% 0/5 — — — 0/5 − 420^(a)    0% 0/5 0% 33.55.0 1/5 ++++ ^(a)Drug was administered 140 mg/kg/injection IV on days 3,5, and 7.

TABLE VI EARLY STAGE MURINE SQUAMOUS LUNG LC-12 (Balb/C female mice,NCI-Kingston) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAY FREEAT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 174RATING   0 +6.5% 0/5 — — — 0/5 −  640^(a)    0% 0/4 14% 12.5 1.7 0/4 ++1391^(b) +1.6% 0/5 20% 15.5 2.1 0/5 +++ ^(a)Drug was administered at 160mg/kg/injection IV on days 4, 6, 8, and 10. ^(b)Drug was administered at107 mg/kg/injection IV on days 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24,26, and 28.

TABLE VII EARLY STAGE HUMAN PROSTATE LNCaP (Balb/C SCID female mice,NCI) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAY FREE ATACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 185 RATING 0 +3.9% — — — — 0/5 − 840^(a) +5.6% 0/5 0 60^(b) 7.2^(b) 4/5 ++++^(a)Drug was administered at 140 mg/kg/injection IV on days 3, 5, 7, 9,10, and 11. ^(b)One mouse.

TABLE VIII UPSTAGED HUMAN BREAST WSU-Br-1 (Balb/C SCID female mice, NCI)TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAY FREE AT ACTIVITY(mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 179 RATING  0 +3.9%— — — — 0/5 − 725^(a) +3.9% 0/5 0% 57 2.9 2/5 ++++ 500^(b)  +14% 0/5 0%39 2.0 2/5 ++++ ^(a)Drug was administered at 145 mg/kg/injection IV ondays 11–13 and 19–20. ^(b)Drug was administered at 100 mg/kg/injectionIV on days 11–13 and 19–20.

TABLE IX EARLY STAGE HUMAN OVARIAN BG-1 (Balb/C SCID female mice, NCI)TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAY FREE AT ACTIVITY(mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 176 RATING  0    0%— — — — 0/5 − 870^(a) −2.0% 0/5 0% 30.5 3.7 1/5 ++++ 600^(b) −3.0% 0/50% 30 3.6 0/5 ++++ ^(a)Drug was administered at 145 mg/kg/injection IVon days 3–5 and 9–11. ^(b)Drug was administered at 100 mg/kg/injectionIV on days 3–5 and 9–11.

TABLE X EARLY STAGE MURINE COLON CARCINOMA-26 ((Balb/C female mice,NCI-Kingston-CRL) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUG DELAYFREE AT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILL DAY 31RATING   0 +1.8%  0/5 — — — 0/5 − 1600^(a) +15% 0/5 34% 15 2.2 0/5 +++1305^(b) +15% 0/5 28% 6 0.9 0/5 + 1740^(c) +17% 0/5 34% 14 2.1 0/5 +++^(a)Drug was administered at 100 mg/kg/injection IV BID on days 1, 3, 5,7, 13, 15, 17, and 21. ^(b)Drug was administered at 145 mg/kg/injectionIV on days 1, 3, 5, 7, 9, 13, 15, 17, and 19. ^(c)Drug was administeredat 145 mg/kg/injection IV on days 1, 2, 3, 7, 8, 13, 14, 16, 18, and20–22.

TABLE XI EARLY STAGE HUMAN SQUAMOUS CELL LUNG CARCINOMA MRI-H165 (Balb/CSCID female mice, NCI) TOTAL % BODY WT GROWTH TUMOR DOSE LOSS AT DRUGDELAY FREE AT ACTIVITY (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOG KILLDAY 55 RATING  0 0% — — — — 0/5 − 600^(a) −2.1%    0/5 0% 31 3.7 0/5++++ ^(a)Drug was administered at 100 mg/kg/injection IV on days 3–5 and10–12.

TABLE XII EARLY STAGE TAXOL RESISTANT HUMAN COLON ADENOCARCINOMA #15/MDR(Balb/C SCID female mice, NCI) % BODY WT GROWTH TUMOR TOTAL LOSS AT DRUGDELAY FREE AT ACTIVITY DOSE (mg/kg) NADIR DEATHS % T/C MASS (DAYS) LOGKILL DAY 38 RATING  0 0% 0/5 — — — 0/5 − 500^(a) +2.1%    0/5 10% 11 1.40/5 ++ ^(a)Drug was administered at 100 mg/kg/injection IV on days 3–5and 13–14.

TABLE XIII EARLY UPSTAGED MURINE COLON ADENOCARCINOMA-38 BY IV, PO, SC,AND IP ROUTES (BDF1 female mice, CRL-Raleigh-NCI) % BODY WT GROWTH TUMORTOTAL LOSS AT DRUG DELAY FREE AT ACTIVITY DOSE (mg/kg) NADIR DEATHS %T/C MASS (DAYS) LOG KILL DAY 161 RATING 0 +6.5% — — — — 0/5 − 450 IV^(a)+8.9% 0/5 0% 18 2.4^(f) 2/5 ++++ 780 PO^(b) +9.7% 0/3 0% Allcures >4.5   3/3 ++++ 450 PO^(c) +6.8% 0/3 0% 22.5 3.0^(f) 2/3 ++++ 450SC^(d) +13.2%  0/2 5.7%   10.5 1.4^(f) 1/2 ++/++++^(g) 450 IP^(e) +8.3%0/2 3.7%   17 2.2^(f) 1/2 +++/++++^(g) ^(a)Drug was administered at 150mg/kg/injection IV on days 5, 7, and 9. ^(b)Drug was administered at 260mg/kg/injection orally on days 5, 7, and 9. ^(c)Drug was administered at150 mg/kg/injection orally on days 5, 7, and 9. ^(d)Drug wasadministered at 150 mg/kg/injection subcutaneously on days 5, 7, and 9.^(e)Drug was administered at 150 mg/kg/injection IP on days 5, 7, and 9.^(f)Log kill of regrowing tumors only (median). ^(g)Activity rating foreach mouse in study. One mouse was cured in each study, representing a“++++” rating. The other activity rating is based on the log killcalculated for the other mouse in the study.

1. A compound of Formula I:

where: X is O; R¹ is hydrogen, halo, C₁–C₆ alkyl, C₁–C₄ alkoxy, C₁–C₄alkylthio, CF₃, OCF₃, SCF₃, (C₁–C₄ alkoxy)carbonyl, nitro, azido,O(SO₂)CH₃, N(CH₃)₂, hydroxy, phenyl, substituted phenyl, pyridinyl,thienyl, furyl, quinolinyl, or triazolyl; R² is hydrogen, halo, cyano,CF₃, C₁–C₆ alkyl, (C₁–C₄ alkoxy)carbonyl, C₁–C₄ alkoxy, phenyl, orquinolinyl; R^(2a) is hydrogen or C₁–C₄ alkoxy; R^(2b) is hydrogen orC₁–C₆ alkyl provided that at least one of R^(2a) and R^(2b) is hydrogen;R³ is hydrogen, halo, C₁–C₆ alkyl, CF₃, or nitro; R^(3a) is hydrogen,halo, or C₁–C₆ alkyl provided that when R^(3a) is C₁–C₆ alkyl, R³ ishydrogen and R⁴ is halo; and R⁴ is halo, C₁–C₆ alkyl, or CF³ providedthat only one of R³ and R⁴ may be C₁–C₆ alkyl and provided that when R⁴is halo or C₁–C₆ alkyl only one of R³ and R^(3a) is hydrogen; or apharmaceutically acceptable base addition salt thereof, provided that:a) when R³ and R⁴ are both chloro and R² is hydrogen, R¹ is bromo, iodo,C₁–C₄ alkoxy, C₁–C₄ alkylthio, CF₃, OCF₃, nitro, azido, O(SO₂)CH₃,N(CH₃)₂, hydroxy, phenyl, substituted phenyl, pyridinyl, thienyl, furyl,or triazolyl; b) when R³ and R⁴ are both chloro and R¹ is hydrogen, R²is bromo, fluoro, CF₃, C₁–C₆ alkyl, C₁–C₄ alkoxy, phenyl, or quinolinyl.2. The compound of claim 1, wherein R², R^(2a), and R^(2b) are hydrogenand R¹ is selected from the group consisting of hydrogen, halo, C₁–C₆alkyl, C₁–C₄ alkoxy, C₁–C₄ alkylthio, CF₃, OCF₃, SCF₃, (C₁–C₄alkoxy)carbonyl, nitro, azido, O(SO₂)CH₃, N(CH₃)₂, hydroxy, phenyl,substituted phenyl, pyridinyl, thienyl, furyl, quinolinyl, andtriazolyl.
 3. The compound of claim 1, wherein the compound is apharmaceutically acceptable base addition salt.
 4. The compound of claim3, wherein the pharmaceutically acceptable base addition salt is asodium salt.
 5. The compound of claim 1 which isN-[2-chloro-4-bromobenzoyl]-4-chlorophenylsulfonamide or a base additionsalt therof.
 6. The compound of claim 1 whichN-[2-methyl-4-chlorobenzoyl]-4-chlorophenylsulfonamide or a baseaddition salt thereof.
 7. The compound of claim 5, wherein the baseaddition salt is a sodium salt.
 8. A method of treating adenocarcinomasof the colon in a mammal comprising administering to a mammal in need ofsuch treatment an oncolytically effective amount of a compound ofFormula II:

where: X is O; R¹ is hydrogen, halo, C₁–C₆ alkyl, C₁–C₄ alkoxy, C₁–C₄alkylthio, CF₃, OCF₃, SCF₃, (C₁–C₄ alkoxy)carbonyl, nitro, azido,O(SO₂)CH₃, N(CH₃)₂, hydroxy, phenyl, substituted phenyl, pyridinyl,thienyl, furyl, quinolinyl, or triazolyl; R² is hydrogen, halo, cyano,CF₃, C₁–C₆ alkyl, (C₁–C₄ alkoxy)carbonyl, C₁–C₄ alkoxy, phenyl, orquinolinyl; R^(2a) is hydrogen or C₁–C₄ alkoxy; R^(2b) is hydrogen orC₁–C₆ alkyl provided that at least one of R^(2a), and R^(2b) ishydrogen; R³ is hydrogen, halo, C₁–C₆ alkyl, CF₃, or nitro; R^(3a) ishydrogen, halo, or C₁–C₆ alkyl provided that when R^(3a) is C₁–C₆ alkyl,R³ is hydrogen and R⁴ is halo; and R⁴ is halo, C₁–C₆ alkyl, or CF₃provided that only one of R³ and R⁴ may be C₁–C₆ alkyl and provided thatwhen R⁴ is halo or C₁–C₆ alkyl only one of R³ and R^(3a) is hydrogen; ora pharmaceutically acceptable base addition salt thereof.
 9. Apharmaceutical formulation comprising a compound of Formula II:

where: X is O; R¹ is hydrogen, halo, C₁–C₆ alkyl, C₁–C₄ alkoxy, C₁–C₄alkylthio, CF₃, OCF₃, SCF₃, (C₁–C₄ alkoxy)carbonyl, nitro, azido,O(SO₂)CH₃, N(CH₃)₂, hydroxy, phenyl, substituted phenyl, pyridinyl,thienyl, furyl, quinolinyl, or triazolyl; R² is hydrogen, halo, cyano,CF₃, C₁–C₆ alkyl, (C₁–C₄ alkoxy)carbonyl, C₁–C₄ alkoxy, phenyl, orquinolinyl; R^(2a) is hydrogen or C₁–C₄ alkoxy; R^(2b) is hydrogen orC₁–C₆ alkyl provided that at least one of R^(2a) and R^(2b) is hydrogen;R³ is hydrogen, halo, C₁–C₆ alkyl, CF₃, or nitro; R^(3a) is hydrogen,halo, or C₁–C₆ alkyl provided that when R^(3a) is C₁–C₆ alkyl, R³ ishydrogen and R⁴ is halo; and R⁴ is halo, C₁–C₆ alkyl, or CF₃ providedthat only one of R³ and R⁴ may be C₁–C₆ alkyl and provided that when R⁴is halo or C₁–C₆ alkyl only one of R³ and R^(3a) is hydrogen; or apharmaceutically acceptable base addition salt thereof, and apharmaceutically acceptable carrier, diluent, or excipient.
 10. Thecompound of claim 2, wherein the compound is a pharmaceuticallyacceptable base addition salt.
 11. The compound of claim 6, wherein thebase addition salt is a sodium salt.